Optimasi Waktu Produksi Endo-β-1,4-D-Xilanase Rekombinan xynbtn63d dalam Saccharomyces Cerevisiae BJ1824

Abstract

Endo–β-1,4-D-xylanase is one of the xylanolitic enzyme that can break the xylosidic bond in xylan to produce xylooligosacharide, these enzyme can be produced by microorganisme such as fungi, bacteria, and Actinomycetes. In this study Endo–β-1,4-D-xylanase were produced by Saccharomyces cerevisiae BJ1824 in pESC-xynBTN63D and pYHM1-xynBTN63D. Optimization of enzyme’s production time can be determined by the growth curve of the isolate using OD measurement every 10 hours for 90 hours, and the activites of Endo–β-1,4-D-xylanase to hydrolized the xylan substrate. The amount of enzyme that can be produced were depend by the amount of S.cerevisiae’s cell, the optimization of incubation time in the production of Endo–β-1,4-D-xylanase was found at 70 hours with the highest total activity 5,003U/mL in pESC-xynBTN63D with a sepesific activity of 7,815U/mg and 4,751 U/mL in pYHM1-xynBTN63D with a sepesific activity of 7,765U/mg. Endo–β-1,4-D-xylanase had an apparent molecular mass of ±30kDa as determined by SDS–PAGE analysis, the electrophoregam shows the thickest protein bond occured at Endo–β-1,4-D-xylanase pESC-xynBTN63D and pYHM1-xynBTN63D that were produced at 70 hours. This analysis indcated the enzyme that were produced at 70 is an optimum enzyme that could degrade oat-spelt xylan as substrate.